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This study was recruited to investigate the role of mitophagy in activating NLRP3 inflammasome in the kidney of uric acid (UA) nephropathy (UAN) rats. This study developed a uric acid nephropathy (UAN) rat model divided into five groups: Negative control (NC), UAN model (M), UAN + autophagy inhibitor (3-MA), UAN + lysosome inhibitor (CQ), and ROS scavenger (N-acetylcysteine, N). H&E staining assessed renal structure, ROS levels were measured with 2, 7-dichlorofluorescin diacetate, and ELISA measured serum markers (creatinine, UA, cystatin C, NGAL, IL-1β, IL-18). Western blot and qRT-PCR evaluated autophagy and inflammation-related protein (LC3 II/I, p62, Pink1, Parkin, NLRP3, Caspase1, IL-1β) expression. NRK-52E cells treated with uric acid and shRNA were analyzed by western blot. Renal injury in UAN rats was aggravated by ROS accumulation, which promoted mitophagy and activated the NLRP3 inflammasome. Eliminating ROS reduced mitophagy, inhibited NLRP3 activation, lowered IL-1β and IL-18 levels, and alleviated renal injury. Notably, inhibiting mitophagy increased ROS accumulation, up-regulated NLRP3, Caspase1, and IL-1β expression, further worsening renal injury. In vitro, uric acid treatment of NRK-52E cells altered autophagy-related protein and pro-inflammatory cytokine levels, highlighting the interplay between mitophagy and inflammation in uric acid nephropathy. Mitophagy influences renal injury in uric acid nephropathy (UAN) by regulating ROS accumulation and NLRP3 inflammasome activation, suggesting that mitophagy may serve as a potential therapeutic target for UAN. [ABSTRACT FROM AUTHOR] more...

N-a-Acetyltransferase 10 protein (Naa10p) is a potential prognostic biomarker and a modulator of several types of cancer. Despite the efforts to elucidate the relationship between Naa10p expression and clinical prognosis, little is known about its expression and role in human oral squamous cell carcinoma (OSCC). In this study, we firstly detected the mRNA and protein levels of Naa10p in 10 paired OSCC tissue samples and found Naa10p was frequently overexpressed in the tumor tissues of patients with OSCC. Further detection by immunohistochemistry was used to examine Naa10p expression in 124 OSCC tumor specimens by tissue microarray (TMA), and a relative high level of Naa10p protein expression was found in 98 out of 124 cases (79.03 %). Additional analyses illustrated that Naa10p expression inversely correlated with clinical stage ( p = 0.047), degree of lymph node status ( p = 0.020), differentiation ( p = 0.022), and recurrence ( p = 0.016) of patients with OSCC. The survival analysis showed that patients with Naa10p-positive expression had a better prognosis for disease-free survival (DFS) or overall survival (OS) than those with Naa10p-negative expression ( p = 0.003 for both). Furthermore, we assessed the effect of Naa10p knockdown on motility of oral cancer cells in vitro, and the results showed that Naa10p inhibit cell wound healing, migration, and invasion. In summary, our study illustrated that the expression of Naa10p had a potential value for predicting the progression of OSCC and prognosis of OSCC patients. [ABSTRACT FROM AUTHOR] more...

Objective To construct chromodomain helicase DNA-bingding protein 4 (CHD4/Mi-2β) truncated recombinant plasmids, induce the expression of GST fusion proteins in E.coli, and purify and identify GST fusion proteins of CHD4/Mi-2β. Methods The gene fragments of chromodomains (CHD4-C), SWI2/SNF2-related ATPase/helicase region (CHD4-H) and DNA-binding domain (CHD4-D) were amplified by PCR and cloned into pGEX-5T vector. After the target genes were sequenced, the plasmids were transformed into E.coli BL21 (DE3) respectively, and induced with 0.8mmol/L IPTG to express fusion proteins. The GST-fusioned recombined proteins were purified with glutathione-agarose resin, and identified by SDS-PAGE and Western blotting. Results Three CHD4 truncated recombinant plasmids were constructed. The recombinant fusion proteins of GST-CHD4-C, GST-CHD4-D and GST-CHD4-H were expressed in E.coli BL21 (DE3) and existed mostly in the form of soluble protein with expected relative molecular weight of 130, 110 and 90ku. The purity of the fusion proteins reached 94.5% after purification by high-affinity glutathione-agarose resin. Conclusion GST-CHD4-C, GST-CHD4-D and GST-CHD4-H have been successfully purified, which lays an experimental foundation for further study of CHD4/Mi-2β in chromatin remodeling. [ABSTRACT FROM AUTHOR] more...

Objective To screen interaction proteins of human PIF1 helicase by yeast-two-hybridization. Methods The proteins interacting with different domains of human PIF1 were screened by using PINT domain (1-180 amino acids) and helicase motifs (167-641 amino acids) of PIF1 helicase as baits respectively to hybridize a HeLa cDNA library. The positive clones were confirmed by one-to-one yeast-two-hybridization, β-galactosidase assay and bioinformatic analysis. Results Totally 17 positive clones interacting with PINT domain of PIF1 were obtained, and 3 positive genes, namely, CCNDBP1, OTUD5 and CAP1, were verified by bioinformatic analysis. However, no positive clone interacting with helicase motifs of PIF1 was obtained. Conclusion The PINT domain of human PIF1 plays a very important role in regulating physiological functions of PIF1 helicase. [ABSTRACT FROM AUTHOR] more...

N-a-Acetyltransferase 10 protein (Naa10p) is a potential prognostic biomarker and a modulator of several types of cancer. Despite the efforts to elucidate the relationship between Naa10p expression and clinical prognosis, little is known about its expression and role in human oral squamous cell carcinoma (OSCC). In this study, we firstly detected the mRNA and protein levels of Naa10p in 10 paired OSCC tissue samples and found Naa10p was frequently overexpressed in the tumor tissues of patients with OSCC. Further detection by immunohistochemistry was used to examine Naa10p expression in 124 OSCC tumor specimens by tissue microarray (TMA), and a relative high level of Naa10p protein expression was found in 98 out of 124 cases (79.03 %). Additional analyses illustrated that Naa10p expression inversely correlated with clinical stage (p = 0.047), degree of lymph node status (p = 0.020), differentiation (p = 0.022), and recurrence (p = 0.016) of patients with OSCC. The survival analysis showed that patients with Naa10p-positive expression had a better prognosis for disease-free survival (DFS) or overall survival (OS) than those with Naa10p-negative expression (p = 0.003 for both). Furthermore, we assessed the effect of Naa10p knockdown on motility of oral cancer cells in vitro, and the results showed that Naa10p inhibit cell wound healing, migration, and invasion. In summary, our study illustrated that the expression of Naa10p had a potential value for predicting the progression of OSCC and prognosis of OSCC patients. more...